Carrier-free column electrophoresis of human lens fluorescent protein derived from water insoluble protein of the lens has been investigated by use of LKB Uniphor column electrophoresis apparatus, in which a sucrose gradient from 10% to 30% in 50 mM Tris-phosphate buffer, pH 7.4 was used for stabilization of separated zone. The voltage applied was 600 volts, and separation time was 18-20 hours. By this method at least three distinct zones were obtained. The fastest moving zone was found to contain the most calcium sensitive protein, indicating that the more negatively charged is the protein, the more sensitive to calcium ion. Best conditions for a large scale preparation of the protein sufficient for the physico-chemical studies are under investigation.